{"files"=>["https://ndownloader.figshare.com/files/2100539"], "description"=>"<p>(A) Immunoblotting analysis was performed to evaluate the interactions between ERα and PHB2 in BIG3- and KPNA (KPNA1, KPNA5, and KPNA6)-depleted MCF-7 cells. MCF-7 cells were treated with siBIG3 and each siKPNA, followed by E2 ± ERAP for 24 h. Then, the nuclear fractions were immunoprecipitated with anti-ERα antibody and were immunoblotted with antibodies against the indicated proteins. The data are expressed the fold increase over E2-treated siBIG3-transfected cells of right and left panels, respectively (set at 1.0). ND: not detected. This experiment was performed using the nuclear fractions used in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0127707#pone.0127707.g003\" target=\"_blank\">Fig 3A</a>; (B) The interaction between ERα and PHB2 released by E2 and ERAP in the nuclear fractions was evaluated. MCF-7 cells depleted of each KPNA were treated with E2 ± ERAP for 24 h, and the nuclear fractions were immunoprecipitated with anti-ERα antibody. The data are expressed the fold increase over E2-treated siEGFP-transfected cells of right and left panels, respectively (set at 1.0). This experiment was performed using the nuclear fractions used in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0127707#pone.0127707.g003\" target=\"_blank\">Fig 3B</a>; (C) The <i>TFF1</i> expression levels following treatment with siBIG3 and siKPNA were evaluated using real-time PCR. The data are expressed as the fold increase over the untreated cells (set at 1.0) and represent the means ± SD of two independent experiments (**<i>P</i><0.01, ***<i>P</i><0.001 in a two-sided Student’s <i>t</i>-test); (D) Immunoblotting analysis was performed to identify the KPNA-binding regions in PHB2. The lysates from COS-7 cells transfected with the indicated HA-PHB2 constructs and FLAG-KPNAs were immunoprecipitated with an anti-FLAG antibody; (E) Immunoblotting analysis was performed to identify the BIG3-binding region in PHB2. The lysates from HEK293T cells transfected with the indicated HA-PHB2 constructs and FLAG-BIG3 were immunoprecipitated with an anti-FLAG antibody. Full-length images of immunoblots are shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0127707#pone.0127707.s006\" target=\"_blank\">S6A–S6D Fig</a>.</p>", "links"=>[], "tags"=>["PHB 2", "BIG 3 Inhibits", "er", "breast cancer cells", "BIG 3 blocks", "overexpressed PHB 2 interacted", "kpna", "BIG 3", "translocation", "PHB 2 tumor suppressor"], "article_id"=>1440637, "categories"=>["Biological Sciences"], "users"=>["Nam-Hee Kim", "Tetsuro Yoshimaru", "Yi-An Chen", "Taisuke Matsuo", "Masato Komatsu", "Yasuo Miyoshi", "Eiji Tanaka", "Mitsunori Sasa", "Kenji Mizuguchi", "Toyomasa Katagiri"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0127707.g005", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_KPNA1_KPNA5_and_KPNA6_induce_E2_dependent_nuclear_translocation_of_PHB2_/1440637", "title"=>"KPNA1, KPNA5, and KPNA6 induce E2-dependent nuclear translocation of PHB2.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2015-06-08 03:46:09"}